Combination of MCL-1 inhibitor with antibody-drug conjugate

Abstract

To provide a more effective method for cancer treatment.SOLUTION: This method includes administering an MCL-1 inhibitor represented by formula (I) and an anti-Trop-2 antibody-drug conjugate. The method may further include administering one or more additional therapeutics selected from chemotherapeutic agents, checkpoint inhibitors, and BTK inhibitors.SELECTED DRAWING: None

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims the benefit of U.S. Provisional Application No. 63 / 209,667, filed Jun. 11, 2021, and U.S. Provisional Application No. 63 / 322,509, filed Mar. 22, 2022. The entire contents of these applications are hereby incorporated by reference in their entirety.

[0002] (Field of the Invention) This application generally relates to a combination therapy of an MCL - 1 inhibitor and an antibody - drug conjugate (ADC) for treating cancer. In particular, the antibody is an anti - Trop - 2 antibody and the drug is an anti - cancer agent.

Background Art

[0003] Apoptosis (programmed cell death) is a process for removing unwanted or potentially dangerous cells from an organism. Avoidance of apoptosis is important for tumor development and sustained growth. Myeloid cell leukemia 1 protein (MCL - 1; also abbreviated as Mcl - 1 or MCL1) is an anti - apoptotic member of the Bcl - 2 family of proteins. MCL - 1 is overexpressed in many cancers. Overexpression of MCL - 1 prevents cancer cells from undergoing apoptosis. Research has shown that MCL - 1 inhibitors can be used to treat various cancers. See, for example: "The MCL1 inhibitor S63845 is tolerable and effective in diverse cancer models", A. Kotschy et al., Nature, 2016(538):477 - 482; "Structure Based Design of "Non-Natural Peptidic Macrocyclic Mcl-1 Inhibitors", J. Johannes et al., ACS Med. Chem. Lett., 2017, 8(2):239-244 & ACS Med. Chem. Lett., 2017, 8(11):1204; "Synergistic action of the MCL-1 inhibitor S63845 with current therapies in preclinical models of triple-negative and HER2-amplified breast cancer", D. Merino et al., Sci. Transl. Med., 2017 Aug. 2, 9(401):eaam7049; "Discovery of Mcl-1-specific inhibitor AZD5991 and preclinical activity in multiple myeloma and acute myeloid leukemia", A. Tron et al., Nature Comm. 2018(9):Article No. 5341; "AMG 176, a Selective MCL1 Inhibitor, Is Effective in Hematologic Cancer Models Alone and in Combination with Established Therapies", S. Caenepeel et al., Cancer Discov., 2018 Dec 8(12):1582-1597; "Discovery of S64315, a Potent and Selective Mcl-1 Inhibitor", Z. Szlavik at al., J. Med. Chem., 2020, 63(22):13762-13795。

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

[0005] In recent years, tumor - related monoclonal antibodies (MAbs) and antibody - drug conjugates (ADCs) containing anti - cancer agents have been developed for the treatment of cancer. For example, sacituzumab govitecan contains the anti - Trop - 2 antibody and SN - 38 disclosed in U.S. Patent No. 7,999,083.

[0006] There remains a need for a more effective method for the treatment of cancer. Summary of the Invention

[0007] In some embodiments, provided herein is a method of treating cancer, the method comprising administering to a human patient in need thereof a therapeutically effective amount of an antibody - drug conjugate and a therapeutically effective amount of an MCL - 1 inhibitor. The antibody-drug conjugate comprises an anti-Trop-2 antibody and an anti-cancer agent; the MCL-1 inhibitor is of formula (I) or a pharmaceutically acceptable salt thereof:

Chemical formula

Brief Description of the Drawings

[0008]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Mode for Carrying Out the Invention

[0009] Definition Unless otherwise required by context, throughout this specification and the claims, the word "comprise" and its variations, such as "comprises" and "comprising", are to be construed in an open and inclusive sense, i.e., "including but not limited to".

[0010] "C u~v ", i.e., a prefix such as (C u ~C v ) indicates that the group following it has u to v carbon atoms, where u and v are integers. For example, "C 1~6 alkyl" indicates that the alkyl group has 1 to 6 carbon atoms.

[0011] A dash ("-") not between two letters or symbols is used to indicate the point of attachment for a substituent. For example, -C(O)NH 2 is attached via a carbon atom. Dashes at the front or end of a chemical group are for convenience, and the chemical group can be shown with or without one or more dashes without losing its normal meaning. Unless chemically or structurally required, the order in which a chemical group is written or named does not indicate or imply directionality.

[0012] The term "substituted" means that one or more hydrogen atoms on a hydrocarbon are substituted with one or more atoms or groups other than hydrogen, provided that the normal valence of the specified carbon atom is not exceeded. A "substituent" is an atom or group that substitutes a hydrogen atom on a hydrocarbon when it is "substituted". Unless otherwise specified, when a group is described as optionally substituted, any substituent of the group is itself unsubstituted.

[0013] References to "about" values or parameters herein include (and describe) embodiments that relate to the value or parameter itself. In certain embodiments, the term "about" includes the indicated amount ±10%. In other embodiments, the term "about" includes the indicated amount ±5%. In certain other embodiments, the term "about" includes the indicated amount ±1%. Also, with respect to that term, "about X" includes the description of "X". Also, the singular forms "a" and "the" include references to the plural unless the context clearly indicates otherwise. Thus, for example, a reference to "a compound" includes a plurality of such compounds, and a reference to "an assay" includes references to one or more assays known to those of skill in the art and their equivalents.

[0014] "Alkyl" refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl has 1 to 20 carbon atoms (i.e., C 1~20 alkyl), 1 to 12 carbon atoms (i.e., C 1~12 alkyl), 1 to 8 carbon atoms (i.e., C 1~8 alkyl), 1 to 6 carbon atoms (i.e., C 1~6 alkyl), 1 to 4 carbon atoms (i.e., C 1~4 alkyl), 1 to 3 carbon atoms (i.e., C 1~3 alkyl), or 1 to 2 carbon atoms (i.e., C 1~2(alkyl). Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl group having a specific number of carbons is named by a chemical name or identified by a molecular formula, all positional isomers having that number of carbons can be included. Thus, for example, "butyl" includes n-butyl (i.e., -(CH 2 ) 3 CH 3 ), sec-butyl (i.e., -CH(CH 3 )CH 2 CH 3 ), isobutyl (i.e., -CH 2 CH(CH 3 ) 2 ) and tert-butyl (i.e., -C(CH 3 ) 3 ), and "propyl" includes n-propyl (i.e., -(CH 2 ) 2 CH 3 ) and isopropyl (i.e., -CH(CH 3 ) 2 ).

[0015] "Aryl" refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or a polycyclic ring system including a fused ring system (e.g., bicyclic or tricyclic). As used herein, aryl has 6 to 20 ring carbon atoms (i.e., C6-20 aryl), 6 to 12 carbocyclic atoms (i.e., C6-12 aryl), or 6 to 10 carbocyclic atoms (i.e., C6-10 aryl). Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, and anthryl. However, aryl does not in any way include, and does not overlap with, heteroaryl as defined below. When one or more aryl groups are fused to a heteroaryl ring, the resulting ring system is heteroaryl.

[0016] "Cycloalkyl" refers to a saturated or partially saturated cyclic alkyl group having a monocyclic or polycyclic ring including a fused ring system, a bridged ring system, and a spiro ring system. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0017] As used herein, "halo" or "halogen" refers to fluoro (-F), chloro (-Cl), bromo (-Br), and iodo (-I).

[0018] As used herein, the term "haloalkyl" refers to alkyl as defined herein, wherein one or more hydrogen atoms of the alkyl are independently substituted with halogen substituents which may be the same or different. For example, C 1~6 Haloalkyl is C 1~6 Alkyl, and C 1~6 One or more of the hydrogen atoms of the alkyl are substituted with halo substituents. Examples of haloalkyl groups include, but are not limited to, fluoromethyl, fluorochloromethyl, difluoromethyl, difluorochloromethyl, trifluoromethyl, 1,1,1-trifluoroethyl, and pentafluoroethyl.

[0019] "Heteroaryl" refers to an aromatic group including a monocyclic, polycyclic, or multiple fused ring having at least one heteroatom in the ring, i.e., one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur, having an aromatic tautomer or resonance structure, wherein nitrogen or sulfur may be oxidized. Thus, the term includes one or more cyclic O, N, S, S(O), S(O) 2and includes a ring having an N-oxide group. This term includes a ring having one or more cyclic C(O) groups. As used herein, heteroaryl has 5 to 20 ring atoms (i.e., 5- to 20-membered heteroaryl), 5 to 12 ring atoms (i.e., 5- to 12-membered heteroaryl), or 5 to 10 ring atoms (i.e., 5- to 10-membered heteroaryl), and independently includes 1 to 5 heteroatoms selected from nitrogen, oxygen, and sulfur, and oxidized forms of the heteroatoms. Examples of heteroaryl groups include pyridin-2(1H)-one, pyridazin-3(2H)-one, pyrimidin-4(3H)-one, quinolin-2(1H)-one, pyrimidinyl, purinyl, pyridyl, pyridazinyl, benzothiazolyl, and pyrazolyl. Heteroaryl does not include aryl as previously defined and does not overlap with aryl.

[0020] The terms “heterocyclyl,” “heterocyclic,” or “heterocyclic ring” refer to a monoradical or diradical saturated or unsaturated group having a single ring or multiple fused rings having one or more heteroatoms selected from nitrogen, sulfur, phosphorus, and / or oxygen within the ring. The heteroatoms within “heterocyclyl” can be oxidized, e.g., -N(O)-, -S(O)-, -S(O) 2 - can be. Heterocyclyl can be monocyclic or polycyclic, and polycyclic can be fused, bridged, or spiro.

[0021] “Isomers” are different compounds having the same molecular formula. Isomers include stereoisomers, enantiomers, and diastereomers.

[0022] “Stereoisomers” refer to compounds having the same atoms connected by the same bonds but having different three-dimensional structures that are not interchangeable. This disclosure contemplates various stereoisomers and mixtures thereof and includes “enantiomers,” which refer to two stereoisomers that are mirror images of each other such that their molecules cannot be superimposed on each other.

[0023] "Tautomer" refers to the proton transfer from one atom of a molecule to another atom of the same molecule. This disclosure includes tautomers of any such compound.

[0024] "Solvate" is formed by the interaction of a solvent and a compound. Solvates of the salts of the compounds described herein are also provided. Hydrates of the compounds described herein are also provided.

[0025] As used herein, the term "prodrug" is a biologically inactive derivative of a drug that is converted to a biologically active parent drug following several chemical or enzymatic pathways upon administration into the human body.

[0026] "Enantiomers" are a pair of stereoisomers that are mirror images that cannot be superimposed on each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The symbol "(±)" is used, as appropriate, to designate a racemic mixture.

[0027] "Diastereomeric isomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other.

[0028] As used herein, "treatment" or "treating" is an approach to obtain a beneficial or desired result. For the purposes of this disclosure, beneficial or desired results include, but are not limited to, alleviation of symptoms and / or reduction of symptoms associated with a disease or condition. In one embodiment, "treatment" or "treating" includes one or more of a) inhibiting a disease or condition (e.g., reducing one or more symptoms resulting from the disease or condition and / or reducing the extent of the disease or condition), b) delaying or halting the onset of one or more symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, delaying the worsening or progression of the disease or condition), and c) alleviating the disease or condition, e.g., regressing clinical symptoms, improving the disease state, delaying the progression of the disease, enhancing the quality of life, and / or extending the survival period.

[0029] As used herein, "prevention" or "preventing" refers to a regimen that protects against the onset of a disease or disorder so that the clinical symptoms of the disease or disorder do not develop. Thus, "prevention" relates to the administration of treatment to a subject before signs of the disease are detectable in the subject. The subject may be an individual at risk of developing a disease or disorder, such as an individual having one or more risk factors known to be associated with the onset or development of the disease or disorder.

[0030] As used herein, the terms "therapeutically effective amount" or "effective amount" refer to an amount effective to induce a desired biological or medical response, including an amount of a drug sufficient to effect such treatment of a disease when administered to a subject for treating the disease. The effective amount varies depending on the particular drug, as well as characteristics of the subject being treated, such as age, weight, etc. The effective amount can include a range of amounts. As understood in the art, the effective amount can be one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired treatment endpoint. The effective amount may be considered in relation to the administration of one or more therapeutic agents, and a single agent may be considered to be administered in an effective amount when used in combination with one or more other agents, whether desirable or beneficial results can be achieved, or are achieved. The appropriate dosage of any co-administered agent may optionally be reduced due to the combined action of the agents (e.g., additive or synergistic effect).

[0031] As used herein, "co - administration" includes administering a unit dose of a drug disclosed herein before or after administration of a unit dose of one or more additional therapeutic agents, e.g., administering the drug disclosed herein within seconds, minutes, or hours of administration of one or more additional therapeutic agents. For example, in some embodiments, a unit dose of a drug of the present disclosure is administered first, and then, within seconds or minutes, a unit dose of one or more additional therapeutic agents is administered. Alternatively, in other embodiments, a unit dose of one or more additional therapeutic agents is administered first, and then, within seconds or minutes, a unit dose of a compound of the present disclosure is administered. In some embodiments, a unit dose of a compound of the present disclosure is administered first, and then, several hours (e.g., 1 - 12 hours) later, a unit dose of one or more additional therapeutic agents is administered. In other embodiments, a unit dose of one or more additional therapeutic agents is administered first, and then, several hours (e.g., 1 - 12 hours) later, a unit dose of a compound of the present disclosure is administered.

[0032] "Administered in combination with" one or more additional therapeutic agents includes simultaneous (concurrent) and sequential or successive administration in any order.

[0033] The term "simultaneously" is used herein to refer to the administration of two or more therapeutic agents where at least a portion of the administrations overlap in time or where the administration of one therapeutic agent occurs within a short period relative to the administration of another therapeutic agent. For example, two or more therapeutic agents are administered at intervals of less than a particular number of minutes.

[0034] The term "sequentially" is used herein to refer to the administration of two or more therapeutic agents where the administration of one or more agents continues after the administration of one or more other agents has been discontinued or where the administration of one or more agents begins before the administration of one or more other agents. For example, two or more therapeutic agents are administered at intervals longer than a particular number of minutes.

[0035] As used herein, "in combination" refers to the administration of one therapeutic modality in addition to another therapeutic modality. Thus, "in combination" refers to the administration of one therapeutic modality before, during, or after the administration of another therapeutic modality to an individual.

[0036] The terms "conjugate" or "antibody-drug conjugate" refer to an antibody that is chemically linked to a second chemical moiety, such as a therapeutic or cytotoxic agent. The term "agent" includes chemical compounds, mixtures of chemical compounds, biological macromolecules, or extracts made from biological materials. In some embodiments, therapeutic or cytotoxic agents include, but are not limited to, pertussis toxin, taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracinedione, mitoxantrone, mitramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, and analogs or homologs thereof. In the context of an immunoassay, a conjugate antibody can be a detectably labeled antibody that is used as a detection antibody.

[0037] "Intravenous administration" is the administration of a substance into a vein or "intravenously". Compared to other routes of administration, the intravenous (IV) route is for delivering fluids and drugs throughout the body It is a faster method. The infusion pump can enable accurate control over the flow rate and total amount of the drug to be delivered. However, if changes in the flow rate do not have serious consequences or the pump is not available, the drip is often simply left to flow by placing the bag above the patient's height and adjusting the speed using a clamp. Alternatively, if the patient requires a high flow rate and the IV access device has a diameter large enough to accommodate it, a rapid infuser can be used. This is either an inflatable cuff placed around the fluid bag to pump the fluid into the patient or a similar electrical device that can also heat the fluid being infused. If the patient requires the drug only at specific times, intermittent infusion without the need for additional fluid is used. This can use the same technology as an intravenous drip (pump or gravity drip), but after the full dose of the drug has been administered, the tube is disconnected from the IV access device. Some drugs are also administered by IV push or bolus. That is, a syringe is connected to the IV access device and the drug is injected directly (slowly if it irritates the vein or causes a rapid effect). When a pharmaceutical is injected into the fluid flow of the IV tube, there must be some means to ensure that it reaches the patient from the tube. Usually, this is achieved by allowing the fluid flow to flow normally, thereby carrying the pharmaceutical into the bloodstream. However, a second fluid injection may be used as a "flush" after injection to more rapidly flush the pharmaceutical into the bloodstream. Thus, in one embodiment, the agent(s) or combination of agents described herein may be administered by IV administration, either alone or in combination with the administration of specific components of a treatment regimen via oral or parenteral routes.

[0038] "Oral administration" is an administration route by which a substance is ingested through the mouth and does not involve direct contact with any of the oral mucosa, such as via a tube, and includes buccal administration, sub labial administration, and sublingual administration, as well as enteral administration and administration through the airway It includes. Typical forms for oral administration of therapeutic agents include the use of tablets or capsules. Thus, in one embodiment, the compound(s) or combination of compounds described herein may be administered orally, either alone or in combination with the administration of specific components of a treatment regimen by the IV or parenteral route.

[0039] Also provided herein are pharmaceutically acceptable salts, hydrates, solvates, tautomers, polymorphs, and prodrugs of the compounds of formula (I) described herein. "Pharmaceutically acceptable" or "physiologically acceptable" refers to compounds, salts, compositions, dosage forms, and other materials suitable for use in human medicine.

[0040] The compounds of formula (I) described herein may be prepared and / or formulated as pharmaceutically acceptable salts. Pharmaceutically acceptable salts are non-toxic salts of the free base form of the compound that have the desired pharmacological activity of the free base. These salts can be derived from inorganic or organic acids or inorganic or organic bases. For example, a compound containing a basic nitrogen can be prepared as a pharmaceutically acceptable salt by contacting the compound with an inorganic or organic acid. Non-limiting examples of pharmaceutically acceptable salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, besylate, xylenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, γ-hydroxybutyrate, glycolate, tartrate, and mandelate. A list of other suitable pharmaceutically acceptable salts can be found in Remington: The Science and Practice of Pharmacy, 21 st Edition, Lippincott Wiliams and Wilkins, Philadelphia, Pa., 2006.

[0041] Non-limiting examples of "pharmaceutically acceptable salts" of the compounds of formula (I) disclosed herein also include alkali metals (e.g., sodium, potassium), alkaline earth metals (e.g., magnesium), ammonium, and NX 4 + (wherein X is C 1 ~C 4It includes salts derived from suitable bases such as (being alkyl). Base addition salts such as sodium salts or potassium salts are also included. MCL-1 inhibitor Compound

[0042] In some embodiments, provided herein is a method of treating cancer, the method comprising administering to a human patient in need thereof a therapeutically effective amount of an antibody-drug conjugate and a therapeutically effective amount of an MCL-1 inhibitor, The antibody-drug conjugate comprises an anti-Trop-2 antibody and an anti-cancer agent; The MCL-1 inhibitor is of formula (I) or a pharmaceutically acceptable salt thereof:

Chemical formula

[0043] In some embodiments of the methods described herein, the MCL-1 inhibitor is a compound of formula (II) or a pharmaceutically acceptable salt thereof: [Chemical formula] Each R 1 、R 2 、R 3 、R 4 、R 5 、and R 6 is provided herein by the methods defined above or elsewhere in this disclosure.

[0044] In some embodiments, the MCL-1 inhibitor is a compound of formula (III), [Chemical formula] or a pharmaceutically acceptable salt thereof:

[0045] In some embodiments, the MCL-1 inhibitor is a compound of formula (I), formula (II) or formula (III) or a pharmaceutically acceptable salt thereof, and R 2 is hydrogen. In some embodiments, R 2 is C 1~3 alkyl. R 2 is methyl.

[0046] In some embodiments, the MCL-1 inhibitor is a compound of formula (I), formula (II) or formula (III) or a pharmaceutically acceptable salt thereof, and R 3 is C 1~3 alkyl. In some embodiments, R 3 is methyl.

[0047] In some embodiments, the MCL-1 inhibitor is a compound of formula (I), formula (II) or formula (III) or a pharmaceutically acceptable salt thereof, and R 4 is hydrogen. In some embodiments, R 5 is C 1~3 alkyl. In some embodiments, R 5 is methyl. In some embodiments, R 6 is Cl.

[0048] In some embodiments, the MCL-1 inhibitor is a compound of formula (I), formula (II) or formula (III) or a pharmaceutically acceptable salt thereof, wherein R 1 is C 1~4 alkyl and C 1~4 optionally substituted with alkoxyl

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0049] In some embodiments, the MCL-1 inhibitor is compound A, N-[(4S,7aR,9aR,10S,11E,14S)-6-chloro-10-methoxy-14-methyl-16-oxide-18-oxo-3’,4’,7,7a,8,9,9a,10,13,14,15,18-dodecahydro-2’H-spiro[1,19-(ethanediylidene)-16λ 4 -cyclobuta[i][1,4]oxazepino[3,4-f][1,2,7]thiadiazacyclohexadecine-4,1’-naphthalene]-16-yl]-3-methoxy-1-methyl-1H-pyrazole-4-carboxamide, and has the following structure:

Chemical formula

[0050] In some embodiments, MCL-1 inhibitors that can be administered include U.S. Patent No. 10,703,733 (Gilead Sciences), AMG-397, AMG-176, PRT-1419, S64315, AZD59991, ABBV-467, International Publication No. WO 2019 / 222112 (Gilead Sciences), WO 2021 / 096860 (GileadCompounds disclosed in, but not limited to, Sciences), No. 2017147410 (Amgen), No. 2019046150 (Amgen), No. 2019036575 (Amgen), No. 2021021259 (Amgen), No. 2019173181 (Amgen), No. 2018183418 (Amgen), No. 2016033486 (Amgen), No. 2018178226 (AstraZeneca), No. 2017182625 (AstraZeneca), No. 2018178227 (AstraZeneca), No. 2020099470 (AstraZeneca), No. 2019211721 (AstraZeneca), No. 2020097577 (Prelude), No. 2020123994 (Prelude), No. 2008104386 (AbbVie), No. 2008104385 (AbbVie), No. 2008131000 (AbbVie), No. 2008130970 (AbbVie), No. 2019035911 (AbbVie), No. 2019035927 (AbbVie), No. 2019035899 (AbbVie), No. 2010049816 (Servier), No. 2020160157 (Servier), No. 2020115183 (Servier), No. 2020099542 (Servier), No. 2015097123 (Servier), No. 2018078064 (Servier), No. 2020254299 (Servier), No. 2018127575 (Servier), No. 2018234433 (Servier), No. 2018015526 (Servier), No. 2016207225 (Servier), No. 2020078875 (Servier), No. 2017125224 (Servier), No. 2020236817 (Servier), No. 2016207226 (Servier), No. 2016207217 (Servier), No. 2016207216 (Servier), and No. 2007147613 (Novartis).

[0051] In some embodiments, the MCL-1 inhibitor is selected from AMG-397, AMG-176, PRT-1419, and S64315. In some embodiments, the MCL-1 inhibitor is AMG-176. In some embodiments, the MCL-1 inhibitor is AMG-397. In some embodiments, the MCL-1 inhibitor is PRT-1419. In some embodiments, the MCL-1 inhibitor is S64315.

[0052] The compounds disclosed herein may contain one or more asymmetric centers and, thus, can give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined as (R)- or (S)- with respect to absolute stereochemistry. The present disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), or (R)- and (S)- isomers may be prepared using a chiral synthon or chiral reagents or resolved using conventional techniques such as chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of a racemic compound (or a racemic compound of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). Similarly, all tautomeric forms are also intended to be included. Formulations

[0053] In the methods provided herein, the MCL-1 inhibitor can be administered as a pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises a compound of formula (I), (II), (III), or Compound A, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises one or more additional therapeutic agents as more fully described hereinafter.

[0054] The pharmaceutical composition containing the MCL-1 inhibitor disclosed in this specification, or a pharmaceutically acceptable salt thereof, may be prepared using one or more pharmaceutically acceptable excipients that can be selected according to normal conventions. "Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, pigment / colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifying agent that is recognized by the US Food and Drug Administration as being acceptable for use in humans or livestock.

[0055] In certain embodiments, the pharmaceutical composition is provided as a solid dosage form including solid oral dosage forms such as tablets. The tablets may contain excipients including lubricants, fillers, binders, and the like. The aqueous composition may be prepared in a sterile form and may generally be isotonic if delivery other than oral administration is intended. All compositions may contain excipients such as those described in Rowe et al, Handbook of Pharmaceutical Excipients, 6 th edition, American Pharmacists Association, 2009. The excipients can include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextrin, hydroxyalkyl cellulose, hydroxyalkyl methyl cellulose, and stearic acid.

[0056] The pharmaceutical compositions disclosed herein are suitable for various routes of administration including oral administration. The compositions may be presented in unit dosage form and may be prepared by any of the methods known in the pharmaceutical art. Such methods include the step of associating the active ingredient (e.g., the compound of the present disclosure or its pharmaceutically acceptable salt) with one or more pharmaceutically acceptable excipients. The compositions may be prepared by uniformly and intimately associating the active ingredient with a liquid excipient or a finely divided solid excipient or both, and then shaping the product, if necessary. The techniques and formulations are generally described in Remington: The Science and Practice of Pharmacy, 21 stIt can be found in Edition, Lippincott Wiliams and Wilkins, Philadelphia, Pa., 2006.

[0057] The compositions described herein that are suitable for oral administration can be presented as discrete units (unit dosage forms) including, but not limited to, capsules, cachets, or tablets, each containing a predetermined amount of the active ingredient. In one embodiment, the pharmaceutical composition is a tablet.

[0058] In some embodiments, the tablets contain Compound A at strengths of 5 mg and 25 mg. In some embodiments, the tablets contain copovidone, lactose monohydrate, microcrystalline cellulose, crospovidone, magnesium stearate, polyvinyl alcohol, titanium dioxide, polyethylene glycol, and talc. Antibody-drug conjugate (ADC)

[0059] In some embodiments, the methods of treating cancer disclosed herein include administering to a human patient in need thereof a therapeutically effective amount of an antibody-drug conjugate and a therapeutically effective amount of an MCL-1 inhibitor. In some embodiments, the antibody-drug conjugate comprises an anti-Trop-2 antibody and an anti-cancer agent.

[0060] In some embodiments, the antibody-drug conjugate is sacituzumab govitecan as disclosed in U.S. Patent No. 7,999,083. In some embodiments, the ADC comprises an antibody-drug conjugate as disclosed in U.S. Patent No. 7,999,083, which is incorporated herein by reference. In some embodiments, sacituzumab govitecan is sacituzumab govitecan-hziy.

[0061] In some embodiments, the anti-Trop-2 antibody-drug conjugate is datopotamab deruxtecan. In some embodiments, anti-Trop-2 antibody-drug conjugates that can be administered include, but are not limited to, the conjugates disclosed in U.S. Patent Nos. 9,850,312, 9,850,312, International Publication Nos. 20240467, and 18036438.

[0062] In some embodiments, the antibody portion of the ADC is an IgG antibody or an antigen-binding antibody fragment. The antibody can be of various isotypes, preferably human IgG1, IgG2, IgG3, or IgG4, more preferably those containing human IgG1 hinge and constant region sequences. The antibody or fragment thereof can be human-mouse chimeric, human-primate chimeric, humanized (human framework and mouse hypervariable (CDR) regions), or fully human antibodies, as well as variants thereof, such as semi-IgG4 antibodies (referred to as "unibodies") (as described in van der Neut Kolfschoten et al. (Science 2007;317:1554-1557)). More preferably, the antibody or fragment thereof may be designed or selected to contain human constant region sequences belonging to a specific allotype, which can result in reduced immunogenicity when the antibody or ADC is administered to a human subject. Preferred allotypes for administration include non-Glml allotypes (nGlml), such as Glm3, Glm3,1, Glm3,2, or Glm3,1,2. More preferably, the allotype is selected from the group consisting of nGlml, Glm3, nGlml,2, and Km3 allotypes.

[0063] In some embodiments, the antibody portion of the ADC is an anti-Trop-2 antibody. In some embodiments, examples of the anti-Trop-2 antibody include TROP2-XPAT (Amunix), BAT-8003 (Bio-Thera Solutions), TROP-2-IR700 (Chiome Bioscience), datopotamab deruxtecan (Daiichi Sankyo, AstraZeneca), GQ-1003 (Genequantum Healthcare, Samsung BioLogics), DAC-002 (Hangzhou DAC Biotech, Shanghai Junshi Biosciences), sacituzumab govitecan (Gilead Sciences), E1-3s (Immunomedics / Gilead, IBC Pharmaceuticals), TROP2-TRACTr (Janux Therapeutics), LIV-2008 (LivTech / Chiome, Yakult Honsha, Shanghai Henlius BioTech), LIV-2008b (LivTech / Chiome), anti-TROP-2a (Oncoxx), anti-TROP-2b (Oncoxx), OXG-64 (Oncoxx), OXS-55 (Oncoxx), humanized anti-Trop2-SN38 antibody conjugate (Shanghai Escugen Biotechnology, TOT Biopharma), anti-Trop2 antibody-CLB-SN-38 conjugate (Shanghai Fudan-Zhangjiang Bio-Pharmaceutical), SKB-264 (Sichuan Kelun Pharmaceutical / Klus Pharma), TROP2-Ab8 (Abmart), Trop2-IgG (Nanjing Medical University (NMU)), 90Y-DTPA-AF650 (Peking University Examples include, but are not limited to, First Hospital, hRS7-CM (SynAffix), 89Zr-DFO-AF650 (University of Wisconsin-Madison), anti-Trop2 antibody (Mediterranea Theranostic, LegoChem Biosciences), and KD-065 (Nanjing KAEDI Biotech).

[0064] Further examples of anti-TROP-2 therapeutic agents include, but are not limited to, E1.BB.3z-92MI (Immunomedics / Gilead), anti-Trop-2 CAR-T (Gilead), Trop-2 CAR-T (Hangzhou Lonzyme Biological Technology), ARB-001 (Arbele), and MT-103 (Myeloid Therapeutics).

[0065] Examples of anti-TROP-2 antibodies include International Publication No. WO2020016662 (Abmart), WO2020249063 (Bio-Thera Solutions), US Patent Application Publication No. US20190048095 (Bio-Thera Solutions), International Publication No. WO2013077458 (LivTech / Chiome), European Patent No. EP20110783675 (Chiome), International Publication No. WO2015098099 (Daiichi Sankyo), WO2017002776 (Daiichi Sankyo), WO2020130125 (Daiichi Sankyo), WO2020240467 (Daiichi Sankyo), US Patent Application Publication No. US2021093730 (Daiichi Sankyo), US Patent No. US9850312 (Daiichi (Sankyo), Chinese Patent No. 112321715 (Biosion), US Patent Application Publication No. 2006193865 (Immunomedics / Gilead), International Publication No. 2011068845 (Immunomedics / Gilead), US Patent Application Publication No. 2016296633 (Immunomedics / Gilead), No. 2017021017 (Immunomedics / Gilead), No. 2017209594 (Immunomedics / Gilead), No. 2017274093 (Immunomedics / Gilead), No. 2018110772 (Immunomedics / Gilead), No. 2018185351 (Immunomedics / Gilead), No. 2018271992 (Immunomedics / Gilead), International Publication No. 2018217227 (Immunomedics / Gilead), US Patent Application Publication No. 2019248917 (Immunomedics / Gilead), Chinese Patent No. 111534585 (Immunomedics / Gilead), US Patent Application Publication No. 2021093730 (Immunomedics / Gilead), No. 2021069343 (Immunomedics / Gilead), US Patent No. 8435539 (Immunomedics / Gilead), No. 8435529 (Immunomedics / Gilead), No. 9492566 (Immunomedics / Gilead), International Publication No. 2003074566 (Gilead), No. 2020257648 (Gilead), US Patent Application Publication No. 2013039861 (Gilead), International Publication No. 2014163684 (Gilead), US Patent Application Publication No. 9427464 (LivTech / Chiome), US Patent No. 10501555 (Abruzzo Theranostic / Oncoxx), International Publication No. 2018036428 (Sichuan Kelun Pharma), No. 2013068946 (Pfizer), No. 2007095749 (Roche), and No. 2020094670 (SynAffix), but not limited thereto.

[0066] Further examples of anti-TROP-2 therapeutic agents include, but are not limited to, those described in International Publication No. WO 2016 / 201300 (Gilead) and Chinese Patent No. CN 108440674 (Hangzhou Lonzyme Biological Technology).

[0067] In some embodiments, the anti-Trop-2 antibody is selected from hRS7, Trop-2-XPAT, and BAT-8003.

[0068] In some embodiments, the anti-Trop-2 antibody is hRS7. In some embodiments, hRS7 is as disclosed in U.S. Patent Nos. 7,238,785, 7,517,964, and 8,084,583, which are incorporated herein by reference. Additional disclosure of hRS7 includes International Patent Publication No. WO 2003 / 074566.

[0069] In some embodiments, the antibody-drug conjugate comprises an anti-Trop-2 antibody and an anti-cancer agent linked by a linker. In some embodiments, examples of the linker include the linker disclosed in U.S. Patent No. 7,999,083. In some embodiments, the linker is CL2A.

[0070] In some embodiments, the drug moiety of the antibody-drug conjugate is a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is selected from doxorubicin (DOX), epirubicin, morpholino doxorubicin (morpholino-DOX), cyanomorpholino-doxorubicin (cyanomorpholino-DOX), 2-pyrrolino-doxorubicin (2-PDOX), CPT, 10-hydroxycamptothecin, SN -38, topotecan, lutetecan, 9-aminocamptothecin, 9-nitrocamptothecin, taxane, geldanamycin, ansamycin, and epothilone. In some embodiments, the chemotherapeutic moiety is SN-38. Formulation

[0071] Suitable routes of administration of the ADC include, but are not limited to, oral, parenteral, subcutaneous, rectal, transmucosal, enteral, intramuscular, intramedullary, intrathecal, direct intraventricular, intravenous, intravitreal, intraperitoneal, intranasal or intraocular injection. Alternatively, for example, the compound may be administered locally rather than systemically by injecting the compound directly into a solid tumor.

[0072] The ADC can be formulated according to known methods for preparing pharmaceutically useful compositions, whereby the ADC is combined in admixture with pharmaceutically suitable excipients. The ADC can be formulated, for example, for intravenous administration by bolus injection, infusion, or continuous infusion. In some embodiments, the antibody is infused over a period of less than about 4 hours. In some embodiments, the antibody is infused over a period of less than about 3 hours. For example, the first 25-50 mg can be infused within 30 minutes, or within 15 minutes, and the remainder infused over the next 2-3 hours. Injectable formulations can be provided in unit dosage form, for example, in ampoules or in multiple-dose containers, with the addition of a preservative. The composition may take the form of, for example, a suspension, solution or emulsion in an oily or aqueous vehicle, and may contain formulating agents such as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, for example, pyrogen-free distilled water, before use. Treatment method

[0073] In some embodiments, the present disclosure provides a combination of an MCL-1 inhibitor and an antibody-drug conjugate for treating cancer. In some embodiments, the antibody-drug conjugate is sacituzumab govitecan and the MCL-1 inhibitor is Compound A.

[0074] In some embodiments, the cancer is a Trop-2 expressing cancer.

[0075] In some embodiments, the cancer is selected from breast cancer, cervical cancer, colorectal cancer, endometrial cancer, epithelial ovarian cancer, esophageal cancer, follicular thyroid cancer, gastric cancer or gastroesophageal junction adenocarcinoma, head and neck cancer, lung cancer, hepatocellular cancer, non-small cell lung cancer, ovarian cancer, prostate cancer, renal cell cancer, small cell lung cancer, urothelial cancer, and urinary tract cancer.

[0076] In some embodiments, the cancer is selected from triple-negative breast cancer (TNBC), HR+ / HER2-breast cancer, urothelial cancer, non-squamous non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), head and neck squamous cell carcinoma (HNSCC), and muscle-invasive bladder cancer (MIBC).

[0077] In some embodiments, the cancer is metastatic. In some embodiments, the cancer is refractory.

[0078] In some embodiments, the cancer is selected from metastatic non-squamous non-small cell lung cancer (mNSCLC), metastatic triple-negative breast cancer (mTNBC), and metastatic soft tissue sarcoma having non-specific histology.

[0079] In some embodiments, the cancer is metastatic non-squamous non-small cell lung cancer (mNSCLC). In some embodiments, the cancer is metastatic triple-negative breast cancer (mTNBC). In some embodiments, the cancer is metastatic soft tissue sarcoma having non-specific histology.

[0080] In some embodiments, the human patient has received at least one other therapy prior to treatment with the combination therapy of an MCL-1 inhibitor and an antibody-drug conjugate. In some embodiments, the human patient has failed other therapies prior to the treatment disclosed herein. In some embodiments, the human patient has failed one chemotherapy.

[0081] In some embodiments, the human patient has failed therapy with an anti-PD1 agent or an anti-PDL1 agent prior to treatment with a combination therapy of an MCL-1 inhibitor and an antibody-drug conjugate.

[0082] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof and the antibody-drug conjugate are administered simultaneously or separately.

[0083] In some embodiments, the MCL-1 inhibitor is Compound A. Generally, the dosage of Compound A administered to humans will vary depending on factors such as the patient's age, weight, height, gender, general medical condition, and medical history. It may be desirable to provide the recipient with a dosage of the antibody-conjugate in the range of about 1 mg / kg to 24 mg / kg as a single intravenous infusion, although lower or higher dosages may be administered depending on the circumstances. For example, a dosage of 1 to 20 mg / kg for a 70 kg patient is 70 to 1,400 mg. The dosage may be repeated, if necessary, for example, once a week for 4 to 10 weeks, once a week for 8 weeks, or once a week for 4 weeks. Also, in maintenance therapy, the frequency may be reduced, if necessary, and administered, for example, at intervals of several months, weekly, or monthly or quarterly over several months. In some embodiments, the dosages include, but are not limited to, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 22 mg / kg, 24 mg / kg, 26 mg / kg, 28 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 55 mg / kg, 65 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 120 mg / kg, 140 mg / kg, 150 mg / kg, 160 mg / kg, 180 mg / kg, 200 mg / kg, 220 mg / kg, 240 mg / kg, 250 mg / kg, 260 mg / kg, 280 mg / kg, 300 mg / kg, 350 mg / kg, 400 mg / kg, 450 mg / kg, 550 mg / kg, 600 mg / kg, 650 mg / kg, 700 mg / kg, 750 mg / kg, and 800 mg / kg. Any amount in the range of 1 to 300 mg / kg may be used. Any amount in the range of 1 to 100 mg / kg may be used.

[0084] In some embodiments, the dosage is administered one or two times per week, multiple times. A minimum dosing schedule of 4 weeks, 8 weeks, 16 weeks, or longer may be used. The dosing schedule may include once-weekly dosing, (i) every week, (ii) every other week, (iii) after 1 week of treatment, a 2-week, 3-week, or 4-week break, (iv) after 2 weeks of treatment, a 1-week, 2-week, 3-week, or 4-week break, (v) after 3 weeks of treatment, a 1-week, 2-week, 3-week, 4-week, or 5-week break, (vi) after 4 weeks of treatment, a 1-week, 2-week, 3-week, 4-week, or 5-week break, (vii) after 5 weeks of treatment, a 1-week, 2-week, 3-week, 4-week, or 5-week break; and (viii) monthly, and may include once or twice weekly dosing in cycles selected from the group consisting of. The cycle may be repeated 4, 6, 8, 10, 12, 16, or 20 or more times.

[0085] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered orally.

[0086] In some embodiments, the amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered at a dosage of about 5 mg / kg, 15 mg / kg, or 50 mg / kg.

[0087] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered at a dosage of about 5 mg / kg.

[0088] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in a 21-day cycle with 2 days of dosing followed by 5 days of drug withdrawal.

[0089] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered up to 105 weeks on days 1, 2, 8, 9, 15, and 16 of each 21-day cycle.

[0090] In some embodiments, the antibody-drug conjugate is administered as an intravenous infusion.

[0091] Generally, the dosage of an antibody-drug conjugate administered to a human will vary depending on factors such as the patient's age, weight, height, gender, general medical condition, and medical history. It may be desirable to provide the recipient with a dosage of the antibody-conjugate in the range of about 1 mg / kg to 24 mg / kg as a single intravenous infusion, although lower or higher dosages may be administered depending on the circumstances. For example, a dosage of 1 - 20 mg / kg for a 70 kg patient is 70 - 1,400 mg. The dosage can be repeated, for example, once a week for 4 - 10 weeks, once a week for 8 weeks, or once a week for 4 weeks as needed. Also, in maintenance therapy, the frequency can be reduced as needed, for example, administered at intervals of several months, or monthly or quarterly over several months. In some embodiments, the dosage includes, but is not limited to, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 22 mg / kg, and 24 mg / kg. Any amount in the range of 1 - 24 mg / kg may be used. In some embodiments, the dosage is administered one or two times a week, multiple times. A minimum dosing schedule of 4 weeks, 8 weeks, 16 weeks or longer may be used. The dosing schedule may include one or two administrations per week in a cycle selected from the group consisting of: (i) weekly, (ii) every other week, (iii) after 1 week of treatment, a rest of 2 weeks, 3 weeks, or 4 weeks, (iv) after 2 weeks of treatment, a rest of 1 week, 2 weeks, 3 weeks, or 4 weeks, (v) after 3 weeks of treatment, a rest of 1 week, 2 weeks, 3 weeks, 4 weeks, or 5 weeks, (vi) after 4 weeks of treatment, a rest of 1 week, 2 weeks, 3 weeks, 4 weeks, or 5 weeks, (vii) after 5 weeks of treatment, a rest of 1 week, 2 weeks, 3 weeks, 4 weeks, or 5 weeks; and (viii) monthly. The cycle may be repeated 4, 6, 8, 10, 12, 16, or 20 or more times.

[0092] In some embodiments, the antibody-drug conjugate may be administered at a frequency of once every two or three weeks, repeating a total of at least three doses. Alternatively, it may be administered twice a week for four to six weeks. If the dose can be reduced to about 200 - 300 mg / m 2 (340 mg for a 1.7 m patient, or 4.9 mg / kg for a 70 kg patient), it may be administered once or even twice a week for four to ten weeks. In some embodiments, the dosing schedule may be shortened, i.e., shortened to two to three months, once every two or three weeks. However, it has been determined that higher doses such as 2 mg / kg once a week or once every two to three weeks can be administered by slow intravenous infusion in repeated dosing cycles. The dosing schedule can optionally be repeated at other intervals, and the dose can be adjusted appropriately for the dose and schedule and administered by various parenteral routes.

[0093] In some embodiments, the antibody-drug conjugate dose is administered on days 1 and 8 of each 21-day cycle.

[0094] In some embodiments, the antibody-drug conjugate is administered at a dose of about 4 mg / kg to about 12 mg / kg. In some embodiments, the antibody-drug conjugate is administered at a dose of about 8 mg / kg to about 12 mg / kg.

[0095] In some embodiments, the antibody-drug conjugate is administered at a dose of about 8 mg / kg, about 10 mg / kg, or about 12 mg / kg.

[0096] In some embodiments, the antibody-conjugate is sacituzumab govitecan. In some embodiments, the dosage of sacituzumab govitecan ranges from about 2 mg / kg to 20 mg / kg as a single intravenous infusion. In some embodiments, the dosage of sacituzumab govitecan ranges from about 6 mg / kg to 10 mg / kg as a single intravenous infusion. In some embodiments, the dosage includes 2 mg / kg, 2.5 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 5.5 mg / kg, 6 mg / kg, 6.5 mg / kg, 6 mg / kg, 6.5 mg / kg, 7 mg / kg, 7.5 mg / kg, 8 mg / kg, 8.5 mg / kg, 9 mg / kg, 9.5 mg / kg, 10 mg / kg, 10.5 mg / kg, 11 mg / kg, 11.5 mg / kg, 12 mg / kg, 12.5 mg / kg, 13 mg / kg, 13.5 mg / kg, 14 mg / kg, 14.5 mg / kg, 15 mg / kg, 15.5 mg / kg, 16 mg / kg, 16.5 mg / kg, 17 mg / kg, 17.5 mg / kg, 18 mg / kg, 18.5 mg / kg, 19 mg / kg, 19.5 mg / kg, and 20 mg / kg. In some embodiments, the dosage of the antibody-conjugate is 7.5 mg / kg.

[0097] In some embodiments, the method further comprises one or more additional therapeutic modalities selected from antibodies, conjugates, gene therapy, chemotherapy, radiation therapy, surgical therapy, BTK inhibitors, and checkpoint inhibitors.

[0098] In some embodiments, the method further comprises radiation therapy.

[0099] In some embodiments, the method further comprises administering one or more additional therapeutic agents. In some embodiments, the additional therapeutic agents are selected from chemotherapeutic agents, checkpoint inhibitors, FLT3 agonists, and BTK inhibitors.

[0100] In some embodiments, the checkpoint inhibitor is selected from an anti-PD-1 agent, an anti-PD-L1 agent, an anti-PD-1 / PD-L1 interaction inhibitor, an anti-CTLA4 agent, and an anti-TIGIT agent.

[0101] In some embodiments, the FLT3 inhibitor is GS-3583. FLT3 agonists also include the agents disclosed in CDX-301 and PCT Publication No. WO 2020 / 263830 A1.

[0102] In some embodiments, the FLT3 agonist is the Fc fusion protein disclosed in International Publication No. WO 2022 / 031876.

[0103] In some embodiments, the present disclosure provides a method for treating cancer. The method includes administering an MCL-1 inhibitor and an antibody-drug conjugate for the treatment of cancer; the method further includes administering one or more additional therapeutic agents, provided that the additional therapeutic agent is not an FLT3 agonist. In some embodiments, the additional therapeutic agent is not an FLT3-Fc fusion protein. In some embodiments, the antibody-drug conjugate is sacituzumab govitecan, the MCL-1 inhibitor is Compound A; the additional therapeutic agent is not an FLT3 agonist. In some embodiments, the additional therapeutic agent is not the FLT3 agonist disclosed in International Publication No. WO 2020 / 263830. In some embodiments, the additional therapeutic agent is not the fusion protein comprising the amino acid sequence of SEQ ID NO: 14 of U.S. Patent No. 11 / 124,582.

[0104] In some embodiments, the method includes administering a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor is selected from an anti-PD-1 agent, an anti-PD-L1 agent, an anti-PD-1 / PD-L1 interaction inhibitor, an anti-CTLA4 agent, and an anti-TIGIT agent. In some embodiments, the checkpoint inhibitor is selected from nivolumab, pembrolizumab, atezolizumab, zimberelimab, and pidilizumab. In some embodiments, the checkpoint inhibitor is selected from ipilimumab, ramucirumab, tremelimumab, durvalumab, avelumab, donbanalimab, and tiragolumab.

[0105] Examples of inhibitors of CTLA4 that can be co-administered include, but are not limited to, ipilimumab, tremelimumab, BMS-986218, AGEN1181, AGEN1884, BMS-986249, MK-1308, REGN-4659, ADU-1604, CS-1002, BCD-145, APL-509, JS-007, BA-3071, ONC-392, AGEN-2041, JHL-1155, KN-044, CG-0161, ATOR-1144, PBI-5D3H5, BPI-002, and the bispecific inhibitors FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), MEDI-5752 (CTLA4 / PD-1), XmAb-20717 (PD-1 / CTLA4), and AK-104 (CTLA4 / PD-1).

[0106] Examples of inhibitors of PD-L1 (CD274) or PD-1 (PDCD1) that can be co-administered include pembrolizumab, nivolumab, semipilumab, pidilizumab, AMP-224, MEDI0680 (AMP-514), spartalizumab, atezolizumab, avelumab, durvalumab, ALN-PDL, BMS-936559, CK-301, PF-06801591, BGB-108, BGB-A317 (tislelizumab), GLS-010 (WBP-3055), AK-103 (HX-008), GB-226, AK-105, CS-1003, HLX-10, MGA-012, BI-754091, PDR-001, AGEN-2034, JS-001 (toripalimab), JNJ-63723283, genolimzumab (CBT-501), LZM-009, BCD-100, LY-3300054, SHR-1201, SHR-1210 (camrelizumab), Sym-021, ABBV-181, PD1-PIK, BAT-1306, RO-6084 (PD-L1 antisense oligonucleotide), STI-1110, GX-P2, RG-7446, mDX-400, (MSB0010718C), CX-072, CBT-502, TSR-042 (dostarlimab), MSB-2311, JTX-4014, BGB-A333, SHR-1316, CS-1001 (WBP-3155), MEDI-0680, enobafolimab (KN-035), KD-033, KY-1003, IBI-308 (sintilimab), HLX-20, KL-A167, STI-A1014, STI-A1015 (IMC-001), BCD-135, FAZ-053, TQB-2450, MDX1105-01, MSB-0010718C, GS-4224, GS-4416, INCB086550, MAX10181, and bispecific inhibitor FPT-155 (CTLA4 / PD-L1 / CD28), PF-06936308 (PD-1 / CTLA4), MGD-013 (PD-1 / LAG-3), FS-118 (LAG-3 / PD-L1), MGD-019 (PD-1 / CTLA4), KN-046 (PD-1 / CTLA4), MEDI-5752 (CTLA4 / PD-1), RO-7121661 (PD-1 / TIM-3), XmAb-20717 (PD-1 / CTLA4),AK-104 (CTLA4 / PD-1), M7824 (PD-L1 / TGFβ-EC domain), CA-170 (PD-L1 / VISTA), CDX-527 (CD27 / PD-L1), LY-3415244 (TIM3 / PDL1), GNS-1480 (epidermal growth factor receptor antagonist; programmed cell death ligand 1 inhibitor), M-7824 (PD L1 / TGFβ bifunctional fusion protein), and INBRX-105 (4-1BB / PDL1) are included, but not limited thereto.

[0107] Examples of PD-1 inhibitors include International Publication No. WO2017112730 (Incyte Corp), WO2017087777 (Incyte Corp), WO2017017624, WO2014151634 (Bristol Myers Squibb Co), WO201317322 (Bristol Myers Squibb Co), WO2018119286 (Incyte Corp), WO2018119266 (Incyte Corp), WO2018119263 (Incyte Corp), WO2018119236 (Incyte Corp), WO2018119221 (Incyte Corp), WO2018118848 (Bristol Myers Squibb Co), WO20161266460 (Bristol Myers Squibb Co), WO2017087678 (Bristol Myers Squibb Co), WO2016149351 (Bristol Myers Squibb Co), WO2015033299 (Aurigene Discovery Technologies Ltd), WO2015179615 (Eisai Co Ltd; Eisai Research Institute), WO2017066227 (Bristol Myers Squibb Co), WO2016142886 (Aurigene Discovery Technologies Ltd), WO2016142852 (Aurigene Discovery Technologies Ltd), WO2016142835 (Aurigene Discovery Technologies Ltd; Individual), WO2016142833 (Aurigene Discovery Technologies Ltd), the same as No. 2018085750 (Bristol Myers Squibb Co), the same as No. 2015033303 (Aurigene Discovery Technologies Ltd), the same as No. 2017205464 (Incyte Corp), the same as No. 2016019232 (3M Co; Individual; Texas A&M University System), the same as No. 2015160641 (Bristol Myers Squibb Co), the same as No. 2017079669 (Incyte Corp), the same as No. 2015033301 (Aurigene Discovery Technologies Ltd), the same as No. 2015034820 (BristolMyers Squibb Co), the same as No. 2018073754 (Aurigene Discovery Technologies Ltd), the same as No. 2016077518 (BristolMyers Squibb Co), the same as No. 2016057624 (BristolMyers Squibb Co), the same as No. 2018044783 (Incyte Corp), the same as No. 2016100608 (BristolMyers Squibb Co), the same as No. 2016100285 (BristolMyers Squibb Co), the same as No. 2016039749 (BristolMyers Squibb Co), the same as No. 2015019284 (Cambridge Enterprise Ltd), the same as No. 2016142894 (Aurigene Discovery Technologies Ltd), the same as No. 2015134605 (BristolMyers Squibb Co), the same as No. 2018051255 (Aurigene Discovery Technologies Ltd), the same as No. 2018051254 (Aurigene Discovery Technologies Ltd), the same as No. 2017222976 (Incyte Corp), the same as No. 2017070089 (Incyte Corp), the same as No. 2018044963 (BristolMyers Squibb Co), the same as No. 2013144704 (Aurigene Discovery Technologies Ltd), the same as No. 2018013789 (Incyte Corp), the same as No. 2017176608 (BristolMyers Squibb Co), the same as No. 2018009505 (BristolMyers Squibb Co), the same as No. 2011161699 (Aurigene Discovery Technologies Ltd), the same as No. 2015119944 (Incyte Corp;Merck Sharp & Dohme Corp), the compound described in Patent No. 2017192961 (Incyte Corp), Patent No. 2017106634 (Incyte Corp), Patent No. 2013132317 (Aurigene Discovery Technologies Ltd), Patent No. 2012168944 (Aurigene Discovery Technologies Ltd), Patent No. 2015036927 (Aurigene; Discovery Technologies Ltd), the compound described in Patent No. 2015044900 (Aurigene Discovery Technologies Ltd), and Patent No. 2018026971 (Arising International) are included, but not limited thereto.

[0108] The PD-1 / PD-L1 inhibitor can be administered in any suitable amount known to those skilled in the art. In some embodiments, the compound of Formula I is administered to a subject in an amount of 0.1 to 1000 mg. Representative amounts of the PD-1 / PD-L1 inhibitor administered to a subject include, but are not limited to, 0.1 to 500 mg, 1 to 100 mg, 1 to 50 mg, or 10 to 50 mg. Other amounts of the PD-1 / PD-L1 inhibitor administered to a subject include, but are not limited to, about 1 mg, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or about 100 mg.

[0109] In some embodiments, the methods described herein further comprise administering an anti-TIGIT antibody such as BMS-986207, RG-6058, domvanalimab, AB308, or AGEN-1307.

[0110] In some embodiments, the methods described herein further comprise administering a BTK (Bruton's tyrosine kinase) inhibitor. Examples of such BTK inhibitors are the compounds disclosed in U.S. Patent No. 7,405,295. Additional examples of BTK inhibitors include, but are not limited to, (S)-6-amino-9-(1-(but-2-ynoyl)pyrrolidin-3-yl)-7-(4-phenoxyphenyl)-7H-purin-8(9H)-one, acalabrutinib (ACP-196), BGB-3111, HM71224, ibrutinib, M-2951, tirabrutinib (ONO-4059), PRN-1008, spebrutinib (CC-292), and TAK-020. In some embodiments, the BTK inhibitor is selected from acalabrutinib, tirabrutinib, zanubrutinib, and PCI-32765.

[0111] In some embodiments, the method further comprises administering a chemotherapeutic agent. In some embodiments, the anti-cancer agent is selected from doxorubicin (DOX), epirubicin , morpholino doxorubicin (morpholino-DOX), cyanomorpholino-doxorubicin (cyanomorpholino-DOX), 2-pyrrolino-doxorubicin (2-PDOX), CPT, 10-hydroxycamptothecin, SN-38, topotecan, lutetecan, 9-aminocamptothecin, 9-nitrocamptothecin, taxane, geldanamycin, ansamycin, and epothilone. In some embodiments, the chemotherapeutic agent is docetaxel. In some embodiments, the chemotherapeutic agent is gemcitabine. In some embodiments, the chemotherapeutic agent is paclitaxel.

[0112] In certain embodiments, when combining the agents of the present disclosure with one or more additional therapeutic agents described herein, the components of the composition are administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more administrations. In certain embodiments, when combining the agents of the present disclosure with one or more additional therapeutic agents described herein, the components of the composition are administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more administrations. In certain embodiments, when combining the agents of the present disclosure with one or more additional therapeutic agents described herein, the components of the composition are administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more administrations.

[0113] Co - administration of the agents disclosed herein and one or more additional therapeutic agents generally refers to co - administration or sequential administration such that a therapeutically effective amount of each agent of the agents disclosed herein and one or more additional therapeutic agents is present in the patient's body.

[0114] Co - administration includes administration of a unit dose of the agent disclosed herein either before or after administration of a unit dose of one or more additional therapeutic agents. The agent disclosed herein can be administered within seconds, minutes, or hours of administration of one or more additional therapeutic agents. For example, in some embodiments, a unit dose of the agent disclosed herein is administered first, followed by administration of a unit dose of one or more additional therapeutic agents within seconds or minutes. Alternatively, in other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed by administration of a unit dose of the agent disclosed herein within seconds or minutes. In some embodiments, a unit dose of the agent disclosed herein is administered first, followed by administration of a unit dose of one or more additional therapeutic agents several hours (e.g., 1 - 12 hours) later. In other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed by administration of a unit dose of the agent disclosed herein several hours (e.g., 1 - 12 hours) later.

[0115] In some embodiments, the present disclosure provides a method for treating or preventing cancer. In certain embodiments, the present disclosure provides a method for treating or preventing cancer, comprising administering to a subject a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer is a blood cancer. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is selected from the group consisting of breast cancer, colorectal cancer, skin cancer, melanoma, ovarian cancer, kidney cancer, small cell lung cancer, non-small cell lung cancer, lymphoma, and leukemia. In some embodiments, the cancer is acute myeloid leukemia.

[0116] In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is bladder cancer.

Examples

[0117] Example 1: In Vitro Synergistic Effect of MCL-1 Inhibitors with SN-38 in TNBC and NSCLC Cell Lines To test the combinability between compound A and SN-38 (a topoisomerase inhibitor), in vitro studies were conducted in a panel of triple-negative breast cancer (TNBC, n = 3) and non-small cell lung cancer (NSCLC, n = 2) cell lines using the Bliss independence model of synergy. Cells were exposed to dose titration matrices of each compound alone and in combination for 72 hours, and then cell viability was determined using the Cell Titer Glo reagent. Strong Bliss synergy scores (> 100) were observed in all cell lines tested. Materials and Methods Cell Culture and Reagents

[0118] The HCC70 (ATCC® CRL-2315), HCC1806 (ATCC® CRL-2335), HCC1187 (ATCC® CRL-2322), NCI-H522 (ATCC® CRL-5810) and H820 (ATCC® HTB-181) cell lines were thawed from liquid nitrogen storage and maintained in RPMI-1640 (Gibco-12633) + 10% HI-FBI (Gibco-16140) + Pen / Strep (100× Gibco-15140) according to ATCC guidelines. Cells were passaged according to ATCC guidelines using 0.25% trypsin / EDTA (1× GIBCO-25200).

[0119] SN-38 and compound A stock (provided by the Gilead sample bank) were directly dispensed into the treatment wells using a D300e Digital Dispenser (vendor) with DMSO (Sigma-D2438) at 0.1% v / v as a vehicle control.

[0120] Cell viability was evaluated using Cell Titer Glo® (Promega #G9241) according to the manufacturer's microplate protocol and luminescence was read on a Biotek Synergy Neo2 plate reader. Cell viability combination assay

[0121] For the synergy matrix assay, cell lines were seeded at 5,000 cells per well in clear-bottom white 96-well plates (Corning #3909) in 100 μL of the recommended cell culture medium. The treatment map consisted of a single-agent dose response for compound A (7 three-fold dilutions + untreated control) or SN-38 (9 three-fold dilutions + untreated control) and a checkerboard matrix of 63 different combinations. The concentration ranges were selected based on the relative sensitivity of each cell line to the compounds. Five plates were used for each combination to generate sufficient replicates to calculate the synergy score with 95% confidence intervals (95% CI).

[0122] The HP D300 dispenser was used to apply the compounds and DMSO vehicle to the cells, which were directly dispensed into the medium according to a checkerboard matrix and incubated at 37 °C / 5% CO 2 / 100% relative humidity for 72 hours, after which cell viability was measured by Cell Titer Glo. Data analysis

[0123] Combined viability data were evaluated for synergy using an Excel template described by Prichard and Shipman {Prichard 1990}. Specifically, the single-component dose curves of SN-38 and Compound A were normalized against the viability (%) on each plate, averaged over five technical replicate experiments, and the theoretical additive kill of the combination was calculated according to the Bliss independence principle. The calculated values were compared with the experimental results generated in a 63-concentration checkerboard. Synergy or antagonism scores were created depending on whether the observed growth inhibition was greater or less than the calculated value, respectively.

[0124] For example, if two compounds (B) and (C) at a given concentration each produced 60% inhibition, their theoretical additive inhibition would be 84% according to the following Bliss independence equation. 60% B +60% C * (100% - 60% B ) = 84% B+C If the experimental result was greater than the calculated value (e.g., 90% inhibition), the difference [6%] was added to the synergy score. If the result was smaller (e.g., 78% inhibition), the difference [6%] was added to the antagonism score.

[0125] These differences were summed across the entire checkerboard (63 wells) to give cumulative synergy and antagonism scores in μM 2Given in units of %, it reflected the 2D surface of the dose-response. The 95% confidence interval adjustment was applied to the synergism and antagonism scores, and each total was compared to the scale based on the original method: scores above 50 were considered moderate synergism, scores above 100 were considered strong synergism, and were highly likely to show a combined effect in vivo {Prichard 1990}.

[0126] The data of the combination assay are shown in three formats. The synergism scores at the 95% confidence interval were averaged from n = 2 assays. Exemplary tabular and graphical percent inhibition matrices for each cell line. Exemplary tabular and graphical synergism matrices at the 95% confidence interval for each cell line.

[0127] To test the combinability between compound A and SN-38 (a topoisomerase inhibitor), in vitro studies were conducted in a panel of TNBC (n = 3) and NSCLC (n = 2) cell lines using the Bliss independence model of synergism. Cells were exposed to the single and combined dose titration matrices of each compound for 72 hours, and then cell viability was determined using the Cell Titer Glo reagent. Strong Bliss synergism scores (>100) were observed in all cell lines tested.

Table 1

Table 2

Table 3

Table 4

Table 5

Table 6

Table 7

Table 8

Table 9

Table 10

Table 11

Table 12

[0128] HCC70 (ATCC® CRL-2315) and HCC1806 (ATCC® CRL-2335) were thawed from liquid nitrogen storage and maintained in RPMI-1640 (Gibco-12633) + 10% HI-FBI (Gibco-16140) + Pen / Strep (100× Gibco-15140) according to ATCC guidelines. MDA-MB-468 (ATCC® HTB-132) was thawed and maintained in DMEM (Gibco-11995) + 10% HI-FBS + Pen / Strep. Cells were passaged according to ATCC guidelines using 0.25% trypsin / EDTA (1× GIBCO-25200).

[0129] Paclitaxel) and compound A stock (provided by the Gilead sample bank) were directly dispensed into the treatment wells using a D300e Digital Dispenser (vendor) with DMSO (Sigma-D2438) used up to 0.1% v / v as a vehicle control.

[0130] Cell viability was evaluated using Cell Titer Glo (trademark) (Promega #G9241) according to the manufacturer's microplate protocol, and luminescence was read using a Synergy Neo2 plate reader.

[0131] Cell lysates for the MSD assay were generated using 1× lysis buffer (10× Cell Signaling CST-9803), 100× protease inhibitor, phosphatase inhibitor I, phosphatase inhibitor II (Meso Scale Discovery Inhibitor Pack R70AA-1, and PMSF (SIGMA catalog number 7626).

[0132] The MCL-BAK and MCL1-BIM dimer assays and the total MCL1 assay were developed by MSD Custom Assay Services and performed using the MSD U-PLEX Development Pack K15227N) and protocol revision "2018 Mar rev 2". GAPDH was determined by MSD using the standard assay K151PWD. All plates were read on an MSD SECTOR Imager 2400 using MSD Read Buffer T (R92TC).

[0133] Protein Simple reagents: EZ Standard Pack1 (PS-ST01EZ: Biotinylated ladder, FL standard, and DTT), peroxide (044-379), Luminal-S (043-311), antibody dilution buffer (042-203), streptavidin HRP (042-414), secondary antibodies: goat anti-rabbit (042-206) and goat anti-mouse (042-205), separation matrix (042-512), stacking matrix (042-513), 10X sample buffer (042-195), wash buffer (042-520), upper running buffer (043-163), lower running buffer (043-162), 384-well plate (040-663), size capillary (55700), Protein Simple Instruments Peggy Sue (trademark) and Sally Sue (trademark). Primary antibodies: MCL1 (CST-94296), FBXW7 (Abcam 109617 and Abcam 171961). Cell viability combination assay

[0134] For the synergy matrix assay, TNBC cell lines were seeded at 10,000 cells per well in clear-bottom white 96-well plates (Corning #3909) in 100 μL of the recommended cell culture medium. After incubating the plates at 37 °C and 100% RH for 20 hours, the cells were exposed to the compounds. The treatment map consisted of single-agent dose responses of compound A (seven 3-fold dilutions from 3 μM to 4 nM + untreated control) or paclitaxel (nine 3-fold dilutions from 3 μM to 0.5 nM + untreated control) and 63 different combinations of checkerboard matrices. Five plates were used for each combination to generate sufficient replicates to calculate the synergy score with 95% confidence intervals (95% CI).

[0135] Paclitaxel and DMSO vehicle were first applied to the cells using an HP D300 dispenser and aliquoted directly into the medium according to a checkerboard matrix. Paclitaxel was incubated for 4 hours, then washed away by medium removal, washed twice with 2×200 μL of pre-warmed complete medium, and finally replaced with 100 μL of pre-warmed complete medium. The cells were then exposed to Compound A in the same checkerboard matrix using the D300 dispenser and incubated for 48 hours prior to measurement of viability by Cell Titer Glo. MSD assay

[0136] For the MSD assay, TNBC cell lines were seeded at 25,000 cells per well in clear-bottom white 96-well plates (Corning #3909) in the recommended cell culture medium. After incubating the plates at 37 °C and 100% RH for 20 hours, the cells were exposed to the compounds. Paclitaxel and DMSO vehicle were first applied to the cells using an HP D300 dispenser and aliquoted directly into the medium. Paclitaxel was incubated for 4 hours, then washed away by medium removal, washed twice with 2×150 μL of pre-warmed complete medium, and finally replaced with 100 μL of pre-warmed complete medium. After an additional 20 hours, the samples were harvested by aspirating the supernatant, and 125 μl of 1× lysis buffer was added to each well. The plates were placed on ice for a short time and transferred to a rocking platform at 4 °C for 20 minutes. The plates were placed on dry ice and snap frozen for 10 minutes and then stored at -80 °C until tested.

[0137] The MCL1 and MCL1 - BAK and MCL1 - BIM dimer assays were performed based on their U - Plex technology using materials and protocols provided by MSD Custom Assay Services. First, plates were prepared using the standard U - PLEX capture antibody coating protocol and then washed three times with 150 μL of MSD wash buffer. 25 μL of sample or standard was added directly to the plates, the plates were sealed, and incubated for 1 hour at room temperature with shaking. The plates were washed three times with 150 μL of wash buffer per well, 50 μL of antibody detection solution was added to each well of the MSD plate, the plates were sealed, and incubated for 1 hour at room temperature with shaking. The plates were washed again three times with 150 μL of wash buffer per well. 150 μL of 2× read buffer was added to each well and the plates were read on an MSD SECTOR Imager 2400.

[0138] The GAPDH assay kit (MSD) was performed according to the manufacturer's protocol using 25 μL of lysate per sample added directly to the plate. The plate was sealed and incubated for 1 hour at room temperature with shaking, then washed three times with 150 μL of wash buffer per well. 25 μL of antibody detection solution was added to each well, the plate was sealed, and incubated for 1 hour at room temperature with shaking, then washed three times with 150 μL of wash buffer per well. Then, 150 μL of 2× read buffer was added to each well and the plate was measured for electrochemiluminescence (ECL) on an MSD SECTOR Imager 2400. Simple Western

[0139] Perform the Simple Western immunoassay in a capillary. Load the sample and reagents onto the assay plate and place it in the Protein Simple Instrument. The cell lysate is automatically loaded into the capillary and separated by size as it moves through the stacking and separation matrices. The separated proteins are then immobilized on the capillary wall via its unique photoactivatable capture chemistry. The target protein is identified using a primary antibody and immunoprobed using an HRP-conjugated secondary antibody and a chemiluminescent substrate. Detect and quantify the resulting chemiluminescent signal.

[0140] Measure FBXW7 expression in cell lines after a 4-hour administration of 1 μM paclitaxel, followed by washing and overnight incubation using Simple Western. Prepare the lysate and dilute it to 0.5 μg / ml in 1× lysis buffer. Run the Simple Western platform on a 384-well plate.

[0141] Markers, internal ladder, and DTT are provided in lyophilized form by Simple Western. Resuspend the reagents as described in the protocol. Add 20 uL of water to the markers. Add 40 μl of water to DTT, mix 20 μl of 10X sample buffer and 20 μl of DTT, and call it Z buffer. Load the markers into 1A. Add 5 μl of the lysate to a 1.7 ml Eppendorf tube. Add 1.2 μl of Z reagent to each sample. Heat the samples at 100°C for 5 minutes, cool, and then spin in a microcentrifuge for 30 seconds. Load the samples into wells A2 - 12. Dilute the primary antibody 1:50 with antibody dilution buffer (6 μl + 294 μl dilution buffer). Load 20 μl of each antibody into lanes 2 - 12. The different antibodies for each column are up to 8 antibodies. Actin diluted 1:300 was used as a loading control. Additionally, load either goat anti - rabbit or goat anti - mouse secondary antibody as needed. Spin the plate at 2.6k for 10 minutes at RT, load it into the device, and run overnight. The target protein is identified using the primary antibody and immunoprobed using the HRP - conjugated secondary antibody and chemiluminescent substrate. Detect and quantify the resulting chemiluminescent signal. Data analysis Regarding Bliss synergy

[0142] Combined survival data were evaluated for synergy using the Excel template described by Prichard and Shipman {Prichard 1990}. Specifically, the single - component dose - response curves of paclitaxel and compound A were normalized against the survival rate (%) on each plate, averaged over five technical replicate experiments, and the theoretical additive killing of the combination was calculated according to the Bliss independence principle. The calculated values were compared with the experimental results generated on a 63 - concentration checkerboard. The synergy or antagonism score was created according to whether the observed growth inhibition was greater or less than the calculated value, respectively.

[0143] For example, if two compounds (B) and (C) at a given concentration each result in 60% inhibition, their theoretical additive inhibition is 84% according to the following Bliss independence equation. 60% B +60% C * (100% - 60% B ) = 84% B+C

[0144] If the experimental result is greater than the calculated value (e.g., 90% inhibition), the difference [6%] is added to the synergistic action score. If the result is smaller (e.g., 78% inhibition), the difference [6%] is added to the antagonistic action score.

[0145] These differences are summed across the entire checkerboard (63 wells) to give cumulative synergistic and antagonistic action scores in μM 2 % units, reflecting the 2D surface of the dose - response. A 95% confidence interval adjustment was applied to the synergistic and antagonistic action scores, and each sum was compared to a scale based on the original method: scores above 50 were considered moderate synergy, scores above 100 were considered strong synergy, and were likely to show a combined effect in vivo {Prichard 1990}.

[0146] The data from the combination assay are presented in three formats. The synergistic action scores at the 95% confidence interval were averaged from n = 2 assays. Exemplary tabular and graphical percent inhibition matrices for each cell line. Exemplary tabular and graphical synergistic action matrices at the 95% confidence interval for each cell line. MSD assay

[0147] For total MCL1, ECL signals were recorded and converted to pg / mL determined via an 8-point standard dose range (0 - 10,000 pg / mL) using calibration controls developed at MSD, and a 4-parameter curve fitting function was used in MSD WorkBench software. Results for MCL1-BAK and MCL1-BIM dimers were converted to pg / mL using the same process, but the standard concentrations were in the range of 0 - 50,000 pg / mL. GAPDH results were recorded and reported as ECL and used to normalize both MCL1 and MCL1 dimers within each sample set. For graphical comparison between analytes of a given cell line, each pg / mL data set was normalized to 100% relative to the vehicle control, with protein-free set to 0%.

[0148] Paclitaxel has been reported to downregulate MCL1 protein levels, at least in part, through upregulation of the MCL1 E3-ligase FBXW7, which targets MCL1 for proteasomal degradation {Wertz 2011}. To confirm this observation, HCC70, MDA-MB-468, and HCC1806 TNBC cell lines were treated with a clinically relevant concentration of paclitaxel (1 μM) for 4 hours (Gianni 1995). After paclitaxel treatment, cells were incubated overnight and protein levels were determined. Comparison of paclitaxel treatment with vehicle control revealed an increase in FBXW7 protein levels and a decrease in MCL1 protein levels (Table 13 and Figure 1). Paclitaxel treatment also resulted in a decrease in the protein levels of MCL1-BAK and MCL1-BIM dimers (Table 13 and Figure 1). These results were observed across all three TNBC cell lines (n = 3 biological replicates).

[0149] The HCC70, MDA-MB-468, and HCC1806 TNBC cell lines were treated with a clinically relevant concentration of paclitaxel (1 μM) for 4 hours (Gianni 1995). Paclitaxel treatment increased the FBXW7 protein level, decreased the MCL1 protein level, and decreased the protein levels of the MCL1-BAK and MCL1-BIM dimers in all three cell lines. HCC70, MDA-MB-468, and HCC1806 were pretreated with increasing doses of paclitaxel (adjusted to C max = 1 μM) for 4 hours to mimic clinical exposure and then exposed to increasing doses of compound A for 72 hours, and Bliss synergy (> 100) was observed.

Table 13

[0150] To determine whether the decrease in MCL1 protein level after paclitaxel treatment led to enhanced sensitivity to compound A, Bliss synergy was used. HCC70, MDA-MB-468, and HCC1806 were pretreated with increasing doses of paclitaxel (adjusted to C max = 1 μM) for 4 hours to mimic clinical exposure and then exposed to increasing doses of compound A. Cells were incubated for 72 hours and cell viability was determined using the CTG reagent. Bliss synergy was observed across all three TNBC cell lines exposed to the combination of compound A and paclitaxel in vitro (Table 15). A Bliss synergy score greater than 100 is considered a strong effect {Prichard 1990}.

Table 14

Table 15

Table 16

Table 17

Table 18

Table 19

Table 20

Table 21-1

Table 21-2

Table 21-3

Table 21-4

Table 21-5

Table 21-6

Table 21-7

Table 21-8

Table 21-9

Table 21-10

Table 21-11

Table 21-12

Table 21-13

Table 21-14

Table 21-15

Table 21-16

Table 21-17

Table 22

Table 23

[0151] The trial was conducted in subjects with advanced solid malignancies to characterize the safety and tolerability of Compound A and Compound A in combination with anticancer therapy. Trial Design

[0152] This is an open-label, multi-center, dose-escalating, and dose-expansion Phase 1a / 1b study to evaluate the safety, tolerability, and PK profile of Compound A, record any DLT (dose-limiting toxicity), and determine the MTD (maximum tolerated dose) and / or RP2D (recommended Phase 2 dose) of Compound A as monotherapy and in combination with anticancer therapy in subjects with advanced solid malignancies. The RP2D is the dose level(s) with acceptable tolerability, exposure, efficacy, and biomarker activity. The study consists of two phases: Phase 1a (dose expansion) followed by Phase 1b (dose expansion). · Phase 1a dose escalation: Part A: Dose escalation of Compound A as monotherapy · Phase 1b dose expansion: Part B: Optional disease-specific cohort of Compound A in combination with anticancer therapy in parallel with Part A Part C: Safety introduction and expansion of Compound A in combination with anticancer therapy after Parts A and B.

[0153] Each part of the study consists of a screening, treatment, and follow-up period. Screening is conducted up to 28 days prior to the first administration of study treatment, during which subject eligibility and baseline characteristics are determined. Part A: Phase 1a dose escalation of Compound A as monotherapy.

[0154] Subjects with advanced solid tumors who have failed or are intolerant to standard therapy or for whom no standard therapy exists are consecutively enrolled to receive Compound A as monotherapy at progressively higher dose levels.

[0155] Dose escalation is conducted using a dose-escalation design based on the 3+3 rule.

[0156] Compound A is administered orally up to 105 weeks on Days 1, 2, 8, 9, 15, and 16 of each 21-day cycle.

[0157] The maximum of six cohorts (i.e., six dose levels) with a target of 3 to 6 subjects each receive compound A at escalating dose levels as monotherapy. The planned starting dose of compound A is 5 mg, and the target doses for the following two cohorts are 15 mg and 50 mg. Subsequent dose levels after the starting dose are determined based on all available clinical data including safety, tolerability, and PK (pharmacokinetics) from the previous cohort, approved by the SRT (Safety Review Team), and potentially up to 300 mg. The increase in dose levels is sub-logarithmic at each subsequent dose escalation.

[0158] The safety and tolerability at each dose level are evaluated by the SRT after all subjects within the cohort have been followed for at least 21 days after the first administration of compound A, or after a subject has had a DLT (dose-limiting toxicity) during the first 21 days of investigational drug administration.

[0159] The first block of each dose consists of 3 subjects. If no subject experiences a DLT during the first 21 days of investigational drug administration, dose escalation is carried out. If 1 subject within the first cohort of 3 subjects experiences a DLT during the first 21 days of investigational drug administration, an additional 3 subjects are enrolled at the same dose level. If no DLT is observed in the additional 3 subjects, dose escalation is carried out. If 2 or more subjects experience a DLT within the first 21 days, dose de-escalation to a lower dose is carried out. The MTD (maximum tolerated dose) is the highest dose level with a subject incidence of DLT of less than 33% during the first 21 days of investigational drug administration.

[0160] Regarding the decision rules to apply, 21 days of treatment with a consistent regimen in any given cohort.

[0161] Throughout the trial, subjects with malignant tumors for which biopsies are available may receive an optional tumor biopsy. These subjects must consent and provide separate specific written consent. Dose Escalation Criteria

[0162] For any given cohort, the sponsor may choose to withhold dosing, select an intermediate dose, or discontinue study enrollment at any time based on a review of preliminary safety and available PK and / or pharmacodynamic data.

[0163] Based on a review of the relevant safety and available PK and / or pharmacodynamic data by the SRT, escalation to higher dose cohorts is only done in the absence of DLT and / or when any pre-specified stopping criteria are met. Dose escalation to subsequent cohorts at a scale greater than semi-logarithmic requires affirmation by at least two-thirds of the SRT.

[0164] Dose-limiting toxicity is defined as the following Compound A-related events that occur within the first 21 days (after the first administration of Compound A). · Grade 4 hematological toxicity lasting more than 21 days. · Consider all Compound A-related Grade 3 non-hematological toxicities lasting more than 7 days and all Compound A-related Grade 4 non-hematological toxicities regardless of duration as DLT. Part B: Optional disease-specific cohorts of Compound A in combination with anti-cancer therapy in parallel with Part A

[0165] During the dose escalation of monotherapy in Part A and prior to the formal dose expansion of the sponsor-nominated and supported disease-specific cohorts featuring combination therapy with Compound A in Part C, the sponsor may choose to nominate and support one or more of the following cohorts aligned with those of Part C for combination therapy with Compound A at any previously evaluated dose in Part A that was considered safe and tolerable by the SRT. · Cohort B1: Metastatic NSCLC (Compound A + docetaxel) · Cohort B2: Metastatic NSCLC (Compound A + sacituzumab govitecan) · Cohort B3: Metastatic TNBC (Compound A + docetaxel) · Cohort B4: Metastatic TNBC (Compound A + sacituzumab govitecan) · Cohort B5: mSTS with non-specific histology (Compound A + docetaxel and gemcitabine)

[0166] Each additional cohort consists of a single such population with a specific combination. Part C: Introduction of the safety and dose escalation of Compound A in combination with other anticancer therapies

[0167] This is an open-label Phase 1b trial using Compound A administered in combination with one or more of the following five disease-specific cohorts nominated and supported by the sponsor after completion of Parts A and B. · Cohort C1: Metastatic NSCLC (Compound A + docetaxel) · Cohort C2: Metastatic NSCLC (Compound A + sacituzumab govitecan) · Cohort C3: Metastatic TNBC (Compound A + docetaxel) · Cohort C4: Metastatic TNBC (Compound A + sacituzumab govitecan) · Cohort C5: mSTS with non-specific histology (Compound A + docetaxel and gemcitabine)

[0168] The RP2D is the dose level(s) with acceptable tolerability, exposure, and biomarker activity.

[0169] The SRT recommends the initial dose of Compound A for combined use for each cohort based on the totality of clinical, safety, PK, and pharmacodynamic data. To ensure that the combination therapy is safe and tolerable in each subject population, at least 3 subjects and a safety introduction group of up to 6 subjects are enrolled.

[0170] The safety introduction adopts the same 3 + 3 design and dose escalation rules as in Part A, and uses the same DLT criteria and DLT evaluation window as in Part A to determine the MTD and / or RP2D. At least 6 subjects need to be treated at a certain dose level before this dose level can be expanded. If the homologous disease-specific cohorts from Part B are investigated at the RP2D, those subjects may be counted and considered against the safety introduction group requirements.

[0171] For cohorts (B1 + C1, B2 + C2, B3 + C3, B4 + C4, B5 + C5), at least 20 subjects at the RP2D, including any subjects from Part B and / or the safety introduction, are enrolled. Cohort C1: Compound A combined with docetaxel in metastatic NSCLC after single-line therapy for metastatic disease

[0172] Cohort C1 evaluates safety and tolerability in subjects with metastatic NSCLC after single-line therapy for metastatic disease, and defines the DLT(s), MTD, and / or RP2D of Compound A combined with docetaxel.

[0173] Compound A is administered on days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0174] Docetaxel is administered as an IV infusion over 1 hour on day 1 of each 21-day cycle at a dose of 75 mg / m 3 of body surface area (BSA), provided that the subject's neutrophil count is acceptable on the dosing day, specifically ≥ 1500 cells / mm 2 The treatment continues for up to 105 weeks unless one or more stopping criteria are met.

[0175] The treatment continues for up to 105 weeks unless one or more stopping criteria are met. Cohort C2: Compound A combined with sacituzumab govitecan in metastatic NSCLC after single-line therapy for metastatic disease

[0176] Cohort C2 evaluates safety and tolerability in subjects with metastatic NSCLC after single-line therapy for metastatic disease and defines the DLT(s), MTD, and / or RP2D of Compound A in combination with sacituzumab govitecan.

[0177] Compound A is administered on Days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0178] Sacituzumab govitecan is administered once weekly as an IV infusion at 10 mg / kg on Days 1 and 8 of each 21-day cycle, provided that the subject's neutrophil count is acceptable on the day of administration, specifically ≥1500 cells / mm 3 or absolute neutrophil count (ANC) ≥1000 cells / mm 3 on Day 8 of any cycle. The first infusion should be administered over 3 hours, and the subject should be observed for signs or symptoms of infusion-related reactions for at least 30 minutes both during and after the infusion. Subsequent infusions, if the previous infusion was acceptable, should be administered over 1 - 2 hours, and the subject should be observed for at least 30 minutes both during and after the infusion.

[0179] Treatment continues for up to 105 weeks unless one or more stopping criteria are met. Cohort C3: Compound A in combination with docetaxel in metastatic TNBC after single-line therapy for metastatic disease.

[0180] Cohort C3 evaluates safety and tolerability in subjects with metastatic TNBC after single-line therapy for metastatic disease and defines the DLT(s), MTD, and / or RP2D of Compound A in combination with docetaxel.

[0181] Compound A is administered on Days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0182] Docetaxel is administered provided that the subject's neutrophil count is acceptable on the day of administration, specifically ≥1500 cells / mm 3administered as an IV infusion over 1 hour on Day 1 of every 21-day cycle at a dose of 75 mg / m 2 of BSA, subject to the condition that it is

[0183] Treatment continues for up to 105 weeks, unless one or more discontinuation criteria are met. Cohort C4: Combination of sacituzumab govitecan and Compound A in metastatic TNBC after single-line therapy for metastatic disease

[0184] Cohort C4 evaluates safety and tolerability in subjects with metastatic TNBC after single-line therapy for metastatic disease, and defines the DLT(s), MTD, and / or RP2D of Compound A in combination with sacituzumab govitecan.

[0185] Compound A is administered on Days 1, 2, 8, 9, 15, and 16 of each 21-day cycle.

[0186] Sacituzumab govitecan is administered once weekly as an IV infusion at 10 mg / kg on Days 1 and 8 of every 21-day cycle, subject to the condition that the subject's neutrophil count is acceptable on the day of administration, specifically ≥1500 cells / mm 3 on Day 1 of any cycle or ANC ≥1000 cells / mm 3 on Day 8 of any cycle. The first infusion should be administered over 3 hours, and the subject is observed for signs or symptoms of infusion-related reactions during and for at least 30 minutes after the infusion. Subsequent infusions, if the previous infusion was acceptable, should be administered over 1 - 2 hours, and the subject is observed during and for at least 30 minutes after the infusion.

[0187] Treatment continues for up to 105 weeks, unless one or more discontinuation criteria are met. Cohort C5: Metastatic soft tissue sarcoma with non-specific histology that has not been previously treated for metastatic disease

[0188] Cohort C5 evaluates safety and tolerability and defines the DLT(s), MTD, and / or RP2D of Compound A in combination with gemcitabine and docetaxel in subjects with previously untreated soft tissue sarcoma.

[0189] Compound A is administered on Days 1, 2, 8, 9, 15, and 16 of a 21-day cycle.

[0190] Gemcitabine is administered as an IV infusion at a fixed dose rate of 900 mg / m2 of BSA over 90 minutes on Days 1 and 8, and docetaxel is administered as 100 mg / m2 BSA IV over 60 minutes on Day 8 of each 21-day cycle.

[0191] Treatment continues for up to 105 weeks unless one or more stopping criteria are met. Duration of treatment

[0192] The investigational drug Compound A is administered up to 105 weeks or until disease progression, unacceptable toxicity, substantial non-compliance with the study procedure or investigational drug, study discontinuation, withdrawal from the study, or the first occurrence of any other reason. Example 8: In vitro combination screening of test Compound A and SN-38 in bladder cancer and prostate cancer.

[0193] This study used a 72-hour proliferation assay to evaluate the combinability of Compound A and SN-38 in a panel of bladder cancer and prostate cancer cell lines. All tests were performed and reported by Horizon Discovery (Cambridge, United Kingdom). The compounds were tested for both single-agent dose response and a 9×9 combination matrix with Compound A. The combination results were ranked using a synergy score metric. Materials and methods

[0194] All cell lines and compounds (except Compound A) were supplied and maintained by Horizon Discovery. Compound A was provided to Horizon Discovery by Gilead Sciences.

Table 24

Table 25

[0195] Cells were thawed from liquid nitrogen storage and grown until they divided with the expected doubling time. Cells were seeded into growth medium in black 384-well tissue culture-treated plates and equilibrated by centrifugation. At the time of treatment, the "time 0" set (untreated) of the assay plates was collected and measured. The treated assay plates were incubated with the compound (in triplicate) for 3 days. After the required treatment time, the Cell Titer Glow (Promega) procedure and data point collection were performed by an automated process. The data were subjected to quality control and analyzed using Horizon-owned software. Data Analysis

[0196] Horizon utilized growth inhibition (GI) as a measure of cell growth. The percentage of GI was calculated by applying the following test and formula. When T < V_0: 100 * (1 - (T - V_0) / V_0) When T ≥ V_0: 100 * (1 - (T - V_0) / (V - V_0)) Where T is the signal measurement value of the test substance, V is the untreated / vehicle-treated control measurement value, and V_0 is the untreated / vehicle control measurement value at time 0 (also colloquially called the T0 plate). This formula is derived from the growth inhibition calculation used in the National Cancer Institute's NCI-60 high-throughput screening.

[0197] A GI reading of 0% indicates no growth inhibition and occurs when the T reading on day 3 is comparable to the V reading for each period. A GI of 100% represents complete growth inhibition (cell division arrest), in which case the cells treated with the compound for 3 days have the same endpoint reading as the T0 control cells. A GI of 200% represents complete death (cytotoxicity) of all cells in the culture well, in which case the T reading on day 3 is lower than the T0 control (near or at a value of 0).

[0198] Monotherapy compound A activity is provided using the GI50 value from the growth inhibition measurement. The maximum response observed is the highest growth inhibition measured with compound A. Both endpoints are reported as the mean of 20 independent results taken from the monotherapy compound A curves across the 20 combinations tested.

[0199] Horizon provided a unique synergy score based on the LOEWE additivity principle to characterize the strength of synergistic interactions for the combinations tested. Horizon also generates synergy results based on three standard models of combination effects: highest single agent (HSA), Bliss independence, and LOEWE additivity as described above. All methods rely on the comparison of the monotherapy dose-response curves to the matrix of combinations. For the purposes of this study report, the inventors focused on the Horizon Synergy Score as it is a comprehensive measure of the combination effect. Any positive score reported indicates a synergistic interaction. The Bliss independence score (related to an in-house method for assessing synergy) is included in the appendix as an alternative presentation of the combination results. Results

[0200] The combination of compound A and SN-38 was tested in a 72-hour in vitro cell viability assay. The combination activity of compound A and SN-38 was determined using multiple synergy models (Table 26). In vitro synergy between compound A and SN-38 was observed in multiple bladder cancer cell line models. [Table 26] Example 9: In vivo activity of compound AF in combination with sacituzumab govitecan against the TNBC cancer cell line model, MDA-MB-468. Materials and methods

[0201] 168 female athymic mice were orthotopically inoculated with 5.0×10^6 MDA-MB-468 cells into the third fat pad (using Matrigel). When the average tumor size reached approximately 200 mm^3, the animals were randomized based on tumor volume. Administration started on Day 0 of the study. [Table 27] Observation during survival

[0202] General observations were performed daily. Body weight, tumor volume, and clinical observations were recorded twice a week. Data analysis

[0203] Descriptive statistics were generated from the study data. The data were evaluated to determine whether parametric or non-parametric analysis was appropriate. For parametric data, analysis of variance (ANOVA) followed by post hoc tests were performed to determine significant differences between treatments, time points, and / or groups. For non-parametric data, appropriate statistical analyses were performed (e.g., Kaplan-Meier survival, Kruskal-Wallis one-way analysis of variance, Mann-Whitney or Wilcoxon rank sum, etc.). Results

[0204] Dosing of 15 or 30 mg / kg (QD (administered every 2 days / 5 days off)) of compound A in combination with 200 mg per mouse (IV QW (administered every 2 days / 1 day off)) of sacituzumab govitecan over 6 weeks against MDA-MB-468 (TNBC) tumors. The non-targeted ADC, h679-SN-38, was tested in combination with compound A as a control. Tumor volume was determined for each treatment group (Figure 7). References Ashkenazi A,Fairbrother WJ,Leverson JD,Souers AJ.From basic apoptosis discoveries to advanced selective BCL-2 family inhibitors.Nat Rev Drug Discov 2017;16(4):273-84。 Gianni L,Kearns CM,Giani A,Capri G,Vigano L,Lacatelli A,et al.Nonlinear pharmacokinetics and metabolism of paclitaxel and its pharmacokinetic / pharmacodynamic relationships in humans.J Clin Oncol 1995;13(1):180-90。 Juin P,Geneste O,Gautier F,Depil S,Campone M.Decoding and unlocking the BCL-2 dependency of cancer cells.Nat Rev Cancer 2013;13(7):455-65。 Prichard MN,Shipman C,Jr.A three-dimensional model to analyze drug-drug interactions.Antiviral Res 1990;14(4-5):181-205。 Ruefli-Brasse A,Reed JC.Therapeutics targeting Bcl-2 in hematological malignancies.Biochem J 2017;474(21):3643-57。 Wertz IE,Kusam S,Lam C,Okamoto T,Sandoval W,Anderson DJ,et al.Sensitivity to antitubulin chemotherapeutics is regulated by MCL1 and FBW7. Nature 2011;471(7336):110-4。 Youle RJ, Strasser A. The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 2008;9(1):47-59。 The present invention provides, for example, the following items. (Item 1) A method for treating cancer, comprising administering to a human patient in need thereof a therapeutically effective amount of an antibody-drug conjugate and a therapeutically effective amount of an MCL-1 inhibitor, wherein the antibody-drug conjugate comprises an anti-Trop-2 antibody and an anti-cancer agent; the MCL-1 inhibitor is of formula (I):

Chemical formula

Chemical formula

Claims

[Claim 1] The invention described in this specification.

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